De Ceuninck F, Willeput J, Corvol M
Institut National de la Santé et de la Recherche Médicale, Paris, France.
J Chromatogr B Biomed Appl. 1995 Apr 21;666(2):203-14. doi: 10.1016/0378-4347(94)00576-q.
In order to purify variant IGF II peptides from human placenta, we have developed a purification procedure combining heparin affinity chromatography and cation-exchange, reversed-phase and size-exclusion HPLC. Two peptides were purified, both having apparent M(r) values of ca. 7300 Da as evaluated by SDS-PAGE. N-Terminal sequencing revealed IGF II and an IGF II variant in which Ser29 was replaced by the tetrapeptide Arg-Leu-Pro-Gly. The final yield of variant IGF II was about eight-fold lower than that of IGF II. Both pure peptides were functionally active as they bound to type I and type II IGF receptors from ovine and human placental membranes, as determined by crosslinking experiments and displacement curve studies.
为了从人胎盘中纯化变异型IGF II肽，我们开发了一种结合肝素亲和色谱以及阳离子交换、反相和尺寸排阻高效液相色谱的纯化方法。纯化出了两种肽，通过SDS-PAGE评估，二者的表观分子量均约为7300 Da。N端测序显示一种为IGF II，另一种是IGF II变异体，其中Ser29被四肽Arg-Leu-Pro-Gly取代。变异型IGF II的最终产量比IGF II低约八倍。交联实验和置换曲线研究表明，这两种纯肽均具有功能活性，因为它们能与来自绵羊和人胎盘膜的I型和II型IGF受体结合。
